Microfluidic based methods to study intercellular communications

Abstract

A microfluidic intercellular communication analysis device includes a coverslip and a Polydimethylsiloxane (PDMS) layer attached to the coverslip, the PDMS layer comprising a plurality of microfluidic channels each having an inlet and an outlet, the plurality of microfluidic channels comprising a donor cell channel structured to receive a donor cell population, a recipient cell channel structured to receive at least a recipient cell population and a matrix channel comprising a diffusion barrier having pores, the donor cell channel and the recipient cell channel each comprising inlets and outlets having an arc angle ranging from 180° to 300°, the arc angle structured to prevent cell aggregation in the inlets, the outlets and/or channel surfaces thereof, wherein upon injecting the donor cell population and the recipient cell population, the target subject is imaged by an imaging device and analyzed for intercellular communication and/or functional characterizations for ensuing intercellular communication effects.

What is claimed is: 
     
         1 . A microfluidic intercellular communication analysis device, comprising:
 a coverslip; and   a Polydimethylsiloxane (PDMS) layer attached to upper surface of the coverslip, the PDMS layer comprising a plurality of microfluidic channels each having an inlet and an outlet, the plurality of microfluidic channels comprising a donor cell channel structured to receive a donor cell population, a recipient cell channel structured to receive at least a recipient cell population and a matrix channel comprising a diffusion barrier having pores, the diffusion barrier being structured to mimic extracellular matrix and conduit a target subject from the donor cell channel to the recipient cell channel through the pores, the donor cell channel and the recipient cell channel each comprising inlets and outlets having an arc angle ranging from 180° to 300°, the arc angle being structured to prevent cell aggregation in the inlets, the outlets and/or channel surfaces of the donor cell channel and the recipient cell channel, wherein upon injection of the donor cell population and the recipient cell population respective cell channels, the target subject is imaged by an imaging device couplable to the microfluidic intercellular communication analysis device and analyzed for intercellular communication thereof, and/or functional characterizations for ensuing intercellular communication effects.   
     
     
         2 . The microfluidic intercellular communication analysis device of  claim 1 , wherein the PDMS layer has a thickness of 3-4 mm such that the magnification of the imaging device required for the target subject is achieved without bending or damaging the coverslip. 
     
     
         3 . The microfluidic intercellular communication analysis device of  claim 1 , wherein the matrix channel is selectively activated by a plasma pulse directed only at the matrix channel using an electrode placed in an outlet of the matrix channel and a tip of a plasma generator placed in an inlet of the matrix channel with inlets and outlets of the donor cell channel and the recipient cell channel being blocked. 
     
     
         4 . The microfluidic intercellular communication analysis device of  claim 1 , wherein the diffusion barrier comprises a porous hydrogel having a pore size appropriate for passing the target subject from the donor cell channel into the recipient cell channel via the matrix channel, the hydrogel comprising a plurality of hydrogels each having different pore sizes so as to allow for size selection of particles that diffuse across each hydrogel and for determining a population of particles that exert observed functional effects, the plurality of hydrogels comprising at least Matrigel and PEGDA gel. 
     
     
         5 . The microfluidic intercellular communication analysis device of  claim 1 , wherein inlets and outlets of the donor cell channel and the recipient cell channel are structured to have a gauge such that a seal forms between surfaces of the inlets and surface of an injection device during cell population injection. 
     
     
         6 . The microfluidic intercellular communication analysis device of  claim 1 , wherein the donor cell channel and the recipient cell channel are structured to have a height that prevents cell aggregation in the inlets, the outlets and/or the channel surfaces thereof, the height ranging from 200 μm to 400 μm. 
     
     
         7 . The microfluidic intercellular communication analysis device of  claim 1 , wherein the PDMS layer is hydrophobized at a temperature ranging from 180° C. to 250° C. for a period ranging from 60 minutes to 70 minutes. 
     
     
         8 . The microfluidic intercellular communication analysis device of  claim 1 , wherein inlet and outlet of the matrix channel each comprise a 16 gauge circumference so as to allow a larger pool of the diffusion barrier comprising a hydrogel to be injected into the matrix channel as compared to hydrogel pools allowed to be injected into matrix channels having inlets and outlets with an 18 gauge circumference. 
     
     
         9 . The microfluidic intercellular communication analysis device of  claim 1 , wherein the matrix channel comprises first and second arrays of transversely spaced-apart matrix ribs, the first array of the transversely spaced-apart matrix ribs is offset from the second array of the transversely spaced-apart matrix ribs by a transverse distance so as to improve diffusion barrier injection into the matrix channel as compared to diffusion barrier injection made into matrix channels having transversely aligned first and second arrays of spaced-apart matrix ribs. 
     
     
         10 . The microfluidic intercellular communication analysis device of  claim 1 , wherein the donor cell channel and the recipient cell channel each comprise inlets having a larger gauge circumference than cell channel inlets structured to accommodate only isolated cells such that the donor cell channel inlet and the recipient cell channel inlet are structured to accommodate target subjects larger than isolated cells, the target subjects comprising tissues or organoids. 
     
     
         11 . The microfluidic intercellular communication analysis device of  claim 1 , wherein the donor or recipient cell population is injected into respective cell channel via a tip of a pipette disposed within respective inlet for a predefined time and volume such that the cell population is transferred from the tip of the pipette to respective cell channel based on gravity, the pipette comprising the cell population. 
     
     
         12 . The microfluidic intercellular communication analysis device of  claim 1 , wherein the donor or recipient cell population is extracted from its respective cell channel via an empty pipette tip disposed within the respective outlet for a predefined volume, the cell population being pushed into the empty pipette tip by injecting cell media into respective inlet via a syringe pump. 
     
     
         13 . The microfluidic intercellular communication analysis device of  claim 1 , wherein the target subject comprises EVs, particles, proteins or nucleic acids, and wherein intercellular communication of the target subject is analyzed for disease progression, utilizing the target subject for delivering medicine, applying therapeutic effects, diagnostic purposes, regenerative medicine, providing immunization against at least infectious diseases or cancer, and any other appropriate investigative research. 
     
     
         14 . A microfluidic intercellular communication analysis system, comprising:
 a microfluidic intercellular communication analysis device including a coverslip and a Polydimethylsiloxane (PDMS) layer attached to upper surface of the coverslip, the PDMS layer comprising a plurality of microfluidic channels each having an inlet and an outlet, the plurality of microfluidic channels comprising a donor cell channel structured to receive a donor cell population, a recipient cell channel structured to receive at least a recipient cell population and a matrix channel comprising a diffusion barrier having pores, the diffusion barrier being structured to mimic extracellular matrix and conduit a target subject from the donor cell channel to the recipient cell channel through the pores, the donor cell channel and the recipient cell channel each comprising inlets and outlets having an arc angle ranging from 180° to 300°, the arc angle being structured to prevent cell aggregation in the inlets, the outlets and/or channel surfaces of the donor cell channel and the recipient cell channel; and   an imaging device couplable to the microfluidic intercellular communication analysis device and structured to automatically and continuously acquire images of the target subject within the plurality of microfluidic channels for a predefined period upon injection of the donor cell population and the recipient cell population into respective cell channels, wherein the acquired images are analyzed for intercellular communication of the target subject, and/or functional characterizations for ensuing intercellular communication effects.   
     
     
         15 . The microfluidic intercellular communication analysis system of  claim 14 , wherein the imaging device comprises a microscope, a camera or other appropriate image capture devices. 
     
     
         16 . A method of performing intercellular communication of a target subject, comprising:
 providing a microfluidic intercellular communication analysis device that includes a coverslip and a Polydimethylsiloxane (PDMS) layer attached to upper surface of the coverslip, the PDMS layer comprising a plurality of microfluidic channels each having an inlet and an outlet, the plurality of microfluidic channels comprising a donor cell channel structured to receive a donor cell population, a recipient cell channel structured to receive at least a recipient cell population and a matrix channel comprising a diffusion barrier having pores, the diffusion barrier being structured to mimic extracellular matrix and conduit the target subject from the donor cell channel to the recipient cell channel through the pores, the donor cell channel and the recipient cell channel each comprising inlets and outlets having an arc angle ranging from 180° to 300°, the arc angle structured to prevent cell aggregation in the inlets, the outlets and/or channel surfaces of the donor cell channel and the recipient cell channel;   injecting the donor cell population including the target subject into the donor cell channel via an inlet of the donor cell channel with an injection device;   injecting the recipient cell population into the recipient cell channel via an inlet of the recipient cell channel with the injection device;   acquiring images of the target subject within the plurality of microfluidic channels via an imaging device over a period; and   analyzing the target subject based at least in part on the acquired images, and/or functional characterizations for ensuing intercellular communication effects.   
     
     
         17 . The method of  claim 16 , wherein the providing a microfluidic intercellular communication analysis device comprises:
 selectively activating the matrix channel by a plasma pulse directed only at the matrix channel using an electrode placed in an outlet of the matrix channel and a tip of a plasma generator placed in an inlet of the matrix channel with inlets and outlets of the donor cell channel and the recipient cell channel being blocked.   
     
     
         18 . The method of  claim 16 , wherein the injecting the donor cell population and the injecting the recipient cell population each comprises:
 injecting into respective cell channel via a tip of a pipette disposed within respective inlet for a predefined time and volume such that the cell population is transferred from the tip of the pipette to respective cell channel based on gravity, the pipette comprising the cell population.   
     
     
         19 . The method of  claim 16 , further comprising:
 extracting at least one of the donor cell population and the recipient cell population.   
     
     
         20 . The method of  claim 19 , wherein the extracting at least one of the donor cell population and the recipient cell population comprises:
 placing an empty pipette tip within respective outlet;   upon placing the empty pipette tip within respective outlet, injecting cell media into respective inlet via a syringe pump; and   receiving the at least one of the donor cell population and the recipient cell population into the empty pipette via the tip for a predefined volume.